小儿止哮平喘颗粒对动物模型止咳化痰平喘作用的实验研究

目的观察小儿止哮平喘颗粒对动物模型的止咳化痰平喘作用。方法以咳嗽、喘憋潜伏期,酚红排泌为指标,观察小儿止哮平喘颗粒对小鼠氨水引咳模型及气管段酚红法、豚鼠整体动物药物引喘模型的影响。结果西药对照组、中浓度组、高浓度组引喘时间及跌倒时间与对照组比较差异均有统计学意义（P＜0．05）;对照组、低及中浓度组酚红浓度与高浓度组比较差异均有统计学意义（P＜0．05）。结论小儿止哮平喘颗粒对动物模型小鼠具有止咳化痰平喘作用。     

The clinical significance of eosinophils in the amniotic fluid in preterm labor

Objective.?White blood cells are not traditionally considered to be normally present in amniotic fluid. This study was conducted after the observation that a patient with preterm labor and intact membranes had eosinophils as a predominant cell in the amniotic fluid, and had an episode of asthma during the index pregnancy. The goal of this study was to determine whether women presenting with preterm labor with eosinophils in the amniotic fluid had a different outcome than those without eosinophils as the predominant white blood cell in the amniotic cavity.

Methods.?This retrospective case–control study included women who presented with preterm labor and intact membranes between 24 and 34 weeks of gestation. Patients underwent an amniocentesis shortly after admission for the assessment of the microbiologic status of the amniotic cavity and/or fetal lung maturity. Amniotic fluid was cultured for aerobic and anaerobic bacteria as well as genital mycoplasmas. Cytologic studies included amniotic fluid white blood cell count and differential, which was performed on cytocentrifuged specimens. Patients with microbial invasion of the amniotic cavity and/or an amniotic fluid white blood cell count >20 cells/mm³ were excluded from the study. Cases were defined as women in whom the differential contained >20% of eosinophils. Controls were selected among women with an amniotic fluid eosinophil count ≤20% and matched for gestational age at amniocentesis. The analysis was conducted with non-parametric statistics.

Results.?The study population consisted of 10 cases and 50 controls. Gestational age and cervical dilatation at admission were similar in both groups. Cases had a lower gestational age at delivery than controls [34.6 weeks, inter-quartile range (IQR) 32–37.3 weeks vs. 38.0 weeks, IQR 35–40 weeks, respectively; p?=?0.018]. The prevalence of preterm delivery ≤35 weeks was higher among patients who had >20% eosinophils than in the control group [50% (5/10) vs. 18% (9/50), respectively; p?=?0.029]. Similar results were observed for delivery at <37 weeks [cases: 70% (7/10) vs. controls: 36% (18/50); p?=?0.046].

Conclusions.?Women with preterm labor and intact membranes who have a large proportion of eosinophils in the amniotic fluid are at an increased risk for spontaneous preterm delivery. These patients may have had an episode of preterm labor related to a type I hypersensitivity reaction.     

慢性脑供血不足的内皮功能变化及药物干预的影响

目的探讨慢性脑供血不足(CCCI)的发病机制和病理过程及药物干预对内皮功能的影响。方法根据日本2000年修订的CCCI诊断标准选择研究对象90例,将其分为基础治疗组(A组)、养血清脑颗粒组(B组)、联合用药组(C组)各30例,同时选正常对照组(D组)27例,用药前后测定血清一氧化氮(NO)、血管内皮素-1(ET-1)水平,检测血管内皮功能,并进行统计学分析。结果A、B、C组治疗前分别与D组比较,NO水平降低,ET-1水平升高,差异有统计学意义(P均<0.01);A组治疗前后比较,血清NO、ET-1水平及内皮依赖性血管舒张功能(EDD)差异均无统计学意义,而B、C 2组治疗后各指标水平均较治疗前有改善(P<0.01),非内皮依赖性血管舒张功能(NEDD)在A、B、C 3组治疗前后无差异;治疗后B、C 2组各指标比较差异均无统计学意义,但分别与A组比较,除NEDD(P>0.05)外其他各指标差异均有统计学意义(P<0.01)。结论CCCI患者存在内皮功能异常,而养血清脑颗粒能够调节血管活性物质水平,改善EDD。     

影响移植后的脂肪颗粒活性因素

目的观察碱性成纤维细胞生长因子（bFGF）、血管内皮细胞生长因子（VEGF）、表皮细胞生长因子（EGF）、胰岛素及自体真皮颗粒对培养及移植的脂肪颗粒活性的影响因素。方法通过对培养中的幼鼠脂肪细胞,分别添加bFGF、VEGF、EGF、胰岛素及自体真皮颗粒,四甲基偶氮唑盐（MTT）法测定各组脂肪细胞活性,分析其作用效果;通过对移植体的大体观察、质量测定、HE染色、微血管数量及免疫组织化学染色等,比较各组添加物对脂肪组织活性的影响。结果除真皮颗粒组及对照组外,各实验组在不同时间点的最佳脂肪细胞活性浓度基本一致,且其细胞活性均高于对照组（P〈0．05）。动物实验中对照组可见较大区域坏死灶、囊样脂肪池。实验组脂肪细胞大小不一,存在大量的小体积脂肪细胞;除胰岛素组外,各实验组移植物残余质量及各组微血管密度均明显高于对照组,但随移植时间的延长差异逐渐缩小。结论bFGF、VEGF、EGF及胰岛素均能不同程度提高脂肪细胞活性,bFGF、VEGF、EGF及自体真皮颗粒均能不同程度地提高脂肪颗粒移植后的移植物质量,改善脂肪组织的血运重建。     

Ubiquitin‐negative,eosinophilic neuronal cytoplasmic inclusions associated with stress granules and autophagy: An immunohistochemical investigation of two cases

Identification of the proteinaceous components of the pathological inclusions is an important step in understanding the associated disease mechanisms. We immunohistochemically examined two previously reported cases with eosinophilic neuronal cytoplasmic inclusions (NCIs) (case 1, Mori et al. Neuropathology 2010; 30: 648–53; case 2, Kojima et al. Acta Pathol Jpn 1990; 40: 785–91) using 67 antibodies against proteins related to cytoskeletal constituents, ubiquitin‐proteasome system, autophagy‐lysosome pathway and stress granule formation. Regional distribution pattern of eosinophilic NCIs in case 1 was substantially different from that in case 2. However, NCIs in both cases were immunonegative for ubiquitin and p62 and were immunopositive for stress granule markers as well as autophagy‐related proteins, including valosin‐containing protein. Considering that eukaryotic stress granules are cleared by autophagy and valosin‐containing protein function, our findings suggest that eosinophilic NCIs in the present two cases may represent the process of autophagic clearance of stress granules.     

In vivo acute treatment with trimethyltin chloride causes neuronal degeneration in the murine olfactory bulb and anterior olfactory nucleus by different cascades in each region

Our earlier study demonstrated that in vivo acute treatment with trimethyltin chloride (TMT) produces severe neuronal damage in the dentate gyrus and cognition impairment in mice. In the present study, we assessed whether TMT was capable of causing neuronal degeneration in the olfactory bulb (OB) and anterior olfactory nucleus (AON) of the mouse brain. An intraperitoneal injection of TMT at the dose of 2.8 mg/kg led to a dramatic increase in the number of degenerating cells, which were reactive with antibody against single-stranded DNA, in the granule cell layer (GCL) of the OB and AON 1 day and 2 days later, respectively. TMT treatment produced a marked translocation of phospho-c-Jun-N-terminal kinase from the cytoplasm to the nucleus in the AON. Expectedly, a marked increase in phospho-c-Jun-positive cells was seen in the AON after the treatment. In addition to the AON, the mitral cell layer of the olfactory bulb showed the presence of phospho-c-Jun-positive cells after the treatment. However, the GCL had no cells positive for either phospho-c-Jun-N-terminal kinase or phospho-c-Jun at any time after the treatment with TMT. Similarly, TMT-induced nuclear translocation of the lysosomal enzyme deoxyribonuclease II was seen in the AON, but not in the GCL. On the other hand, TMT elicited the expression of activated caspase 3 in the GCL but not in the AON. Taken together, our results suggest that TMT is capable of causing neuronal degeneration in the murine OB and AON through different cascades in the two structures.     

Cannabinoid agonist WIN55,212-2 induces apoptosis in cerebellar granule cells via activation of the CB1 receptor and downregulation of bcl-xL gene expression

The endogenous cannabinoid system is involved in the regulation of a number of physiologic effects in both the central and peripheral nervous systems. Its role in the control of neuronal cell proliferation has attracted major attention because of its potential implications for new therapeutic strategies. In the present study, we demonstrated that treatment of cultured cerebellar granule cells with the synthetic cannabinoid WIN55,212-2, causes cell-body and nuclear shrinkage, which are hallmarks of neuronal apoptosis, as well as concentration-dependent decrease in cell viability. Staining with the fluorescent nuclear dye, Hoechst 33258, revealed apoptosis in 27.1% and 58.5% of cells exposed to 1 and 10 microM of WIN55,212-2, respectively (P < 0.01 and P < 0.001 vs. control respectively) after 36 hr. After 24 hr of exposure to WIN55,212-2, mRNA levels for the anti-apoptotic gene bcl-xL were reduced to 45.6% of those found in control (P < 0.01). These effects were completely reverted when cells were exposed to the synthetic cannabinoid in the presence of the specific CB1-receptor antagonist, SR141716A (1 microM). Moreover, the pro-apoptotic effect of 10 microM WIN55,212-2 could be reduced by the addition to the incubation medium of a cell-permeant inhibitor of caspase-1 (50 nM). Finally, WIN55,212-2 significantly increased caspase-1 activity after 24 hr. These findings show that the activation of CB1 receptors on cerebellar granule cells induces apoptotic cell death, which is associated with downregulation of the anti-apoptotic gene, bcl-xL, and at least in part, activation of caspase-1.     

Pronounced individual variation in the response to the stimulatory action of exercise on immature hippocampal neurons

In the adult hippocampus, neurogenesis is influenced both by external stimuli, such as physical exercise, and by intrinsic conditions like age and disease. However, the way in which many of these external and internal cues interact in this process remains poorly understood. We have used a new, more precise, stereological cell counting method that involves confocal microscopy to analyze the effects of exercise on adult neurogenesis in the mouse. We found that treadmill exercise increases the number of differentiating neurons (doublecortin/calretinin cells) in the granule cell layer of the mouse hippocampus in a manner that is directly related to the size of the mature granule cell population. More immature neurons were found after exercise in animals that had a larger dentate gyrus (DG), while no changes were observed in those with a smaller DG. This differential response to physical exercise suggests that the pre-existing neuronal population regulates the neurogenic response in the DG to external stimuli. These data raise the possibility of anticipating an individuals' response to therapeutic interventions (like exercise) aimed at augmenting dentate neurogenesis and alleviating or preventing cognitive decline.     

Hippocampal hypocellularity in the Ts65Dn mouse originates early in development

Ts65Dn, a well-characterized animal model for Down syndrome, has three copies of the distal end of mouse chromosome 16 and therefore has segmental trisomy for orthologs for nearly half of the genes located on human chromosome 21. Ts65Dn mice have learning and memory impairments, especially in tasks involving the hippocampus. Previous studies have shown that older adult Ts65Dn mice have structural abnormalities in the hippocampus including fewer granule cells in dentate gyrus and more pyramidal cells in the CA3 subfield of cornus ammonis. However, it is not clear whether those changes are secondary to the age-related neurodegeneration of the basal forebrain cholinergic neurons that project to the hippocampus or if they originate earlier during hippocampal development. To address this question, we performed a quantitative study of the hippocampal volume and the numbers of granule cell and pyramidal neurons in young (postnatal day 6, P6) and adult (3-month-old) mice using the optical fractionator method. At P6, Ts65Dn mice had 20% fewer granule cells in dentate gyrus than did euploid littermates. Similarly, compared to euploid, P6 trisomic mice showed an 18% reduction in mitotic cells in the granule cell layer and the hilus, where granule cell precursors divide to generate the internal granule cell layer. Granule cell hypocellularity persists in 3-month-old Ts65Dn mice before the onset of cholinergic atrophy. The hypocellularity seen in the trisomic adult hippocampus originates early in development and may contribute to specific cognitive deficits in these mice.